Encapsulation, characterisation and in vitro useability of cellulase purified with three-phase partitioning technique

In this thesis, the purification of Trichoderma reesei cellulase which is frequently used in commercial arena by three-phase partitioning (TPP) method and encapsulation in calcium alginate gel was studied. These two basic experiments were optimized with all variables, and finally optimum conditions for both purification and encapsulation studies were determined. Ideal TPP condition for the purification of T. reesei cellulase was determined as; 50% (w/v) ammonium sulfate saturation, 1:1 (v/v) enzyme: t-butanol ratio, pH 4.5 and 0.1 mg of the protein. Under optimum conditions, the cellulase enzyme was successfully purified with 113% activity yield and 1.42 purification fold. As a result of the encapsulation studies, optimum immobilization conditions for commercial cellulase enzyme were determined as; 0.5 mg enzyme (protein amount) and 2% (w/v) alginate, 0.5% (w/v) chitosan in 0.15 M CaCl2, 3 hours, 2.0% (v/v) glutaraldehyde 2 hour cross-linking time. The cellulase enzyme was successfully immobilized with 53% activity yield with this optimum conditions. The cellulase enzyme, which was concentrated in the middle phase of TPP, could be immobilized with 10% activity yield under these optimal conditions.